• Title/Summary/Keyword: red carotenoprotein

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Purification and Characterization of the Red Carotenoprotein from the Skin of Ascidian, Halocynthia roretzi 1. Purification and Characterization of the Caritenopritein (멍게껍질로부터 분리제정한 적색 Carotenoprotein의 특성 1. Carotenoprotein 의 정제 및 특성)

  • Kang, Ok-Ju;Suh, Myung-Ja;Lee, An-Jong;Kim, Se-Kwon
    • Journal of Life Science
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    • v.5 no.4
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    • pp.170-180
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    • 1995
  • A carotennnoprotein from the skin of Ascidian(Halocynthia roretzi) was extracted by Triton X-100 and purified by ammonium sulfate fraction, SephadexG-200 charomatography and DEAE-cellulose ion exchange chromatography. The carotenoprotein was redwith broad $\lambda$$_{max}$ between 495, 467 and 318nm. The red carotenoprotein had an approximate molecular weight of 326KDa(gel filtration). SDS-PAGE indicated the presence of two polypeptodes of 84.1KDa and 74.4KDa, with different mobility in polyacrylamide gel electrophoresis. In the presence of denaturing agents such as organic solvent aand extreme pH, the red complex readily disociates to liberate the yellow carotenoid($\lambda$$_{max}$ 452nm) and a colourless apoprotein. The amino acid composition of carotenoprotein were mainly threonine(15.2%), aspartic acid(12.2%), glutamic acid(11.9%) and serine(9.6%), while proline was not found. The carotenoprotein consisted of lipids as structure units. Its major fatty acids composion were C$_{18:1}$, C$_{16:1}$, and C$_{16:0}$. The monounsaturated fatty acids(41.5%) contained abundant content compared to other fatty aacids(polyunsaturated fatty acids 37.4%, saturated fatty acids 20.6%).

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Purification and Characterization of the Red Carotenoprotein from the Skin of Ascidian, Halocynthia roretzi 2. Chemical Structure of Carotenoid (멍게 껍질로부터 분리정제한 적색 Carotenoprotein의 특성 2. Carotenoid의 화학구조)

  • Kang, Ok-Ju;Suh, Myung-Ja;Lee, An-Jong;Kim, Se-Kwon
    • Journal of Life Science
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    • v.5 no.4
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    • pp.181-189
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    • 1995
  • Carotenoids liberated from the red carotenoprotein from the skin of ascidian(Halocynthia roretzi)were developed on TLC, and two bands were obtained with different Rfvalues. Their chemical reactivities and spectroscopic properties were studied and elucidated as astaxanthin diester and astaxanthin monoester. After saponification of carotenoid ester, GC analysis was performed. The carotenoid ester contained oleic acid, palmitic acid and palmitoleic acids as 50% of total amount, and had higher content unsaturated fatty acid including eicosapentanoic acid than saturated fatty acid.

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Purification and Characterization of the Red Carotenoprotein from the Muscle of Blue Mussel, Mytilus edulis (진주담치 근육중의 적색 Carotenoprotein의 정제 및 특성)

  • YANG Huyn-Pil;LEE An-Jong;KIM Yong-Tae;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.482-494
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    • 1994
  • Most of carotenoprotein complexes have been extracted by using buffered solutions. However, in this study carotenoprotein from the muscle of Blue mussel(Mytilus edulis) was extracted by a detergent such as Triton X-100. It was purified and characterized by $20\%$ (w/v) $(NH_4)_2SO_4$, DEAE-cellulose ion exchange and Sephacryl S-300 gel filtration. The carotenoprotein(${\lambda}_{max}=462nm$) had an approximate M. W. of 372KDa(gel filtration). SDS-PAGE analysis of the carotenoprotein indicated the presence of four polypeptides of 60KDa($23.70\%$), 46.9KDa($9.14\%$), 26KDa($49.14\%$) and 13KDa($18.02\%$). Carotenoprotein denaturated by treatment with SDS to a final concentration of $0.2\%$ (w/v) caused a hypsochromic shift of ${\lambda}_{max}$ from 462nm to 456nm. The carotenoprotein contained lipids as structure units. The amino acid composition of the carotenoprotein contained large essential amino acid amounts of $62.8\%$, and the content of threonine($35.9\%$) was higher than other amino acids, but histidine, methionine and proline were not present. In the carotenoprotein, the major fatty acids were $C_{16:4},\;C_{16:0},\;C_{20:5}\;and\;C_{22:6}$. The percentages of polyunsaturated fatty acids($62.4\%$) were higher compared to other fatty acids(saturated fatty acids $19.6\%$, monounsaturated fatty acids $18.0\%$). Carotenoid was extracted from the carotenoprotein by acetone and it was separated into five different components by preparative TLC(benzene:petroleum ether:acetone=69:17:14). The major components of carotenoid were mytiloxanthin($74.79\%$) and 3,4,3'- trihydroxy-7',8'-didehydro-${\beta}$-carotene($18.26\%$), and they were at least presented as prosthetic groups of carotenoprotein.

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